LTP is triggered by Ca^* entry through the NMDAR;subsequently Ca^* activates calmodulin (CaM), which then activates CaMKIl. Despite extensive studies demonstrating the pivotal role of CaMKIl in LTP and memory, the mechanisms of its activation in living cells is not known. The goal ofthe proposed work is to understand these mechanisms in quantitative detail. This requires methods to measure biochemical events in single spines near the limit of optical resolution and a sophisticated modeling framework for simulating these reactions. Because the experimental and computational methods were not previously available, this will be the first attempt to account for the measured activation of an enzyme in a living cell.
Aim 1. Measurements will be made of critical quantitative properties of the system, including free CaMKIl, CaM, Ng and CaMKIl. This will be done used calibrated optical methods.
Aim 2. To model CaM activation requires information about the spatial/temporal gradients of Ca^* in spines. 2-photon uncaging of glutamate will be used to activate NMDARs in a controlled way;the resulting Ca^* elevation in the bulk spine cytoplasm will be measured.
Aim 3. Using fluorescence lifetime methodology (FLIM)), the time course of CaMKIl activation in single spines will be measured. Computer simulations will then be used to predict how the elevation of Ca^*, as determined in Aim 2, leads to CaMKIl activation. The parameters determined in Aim 1 are needed for this calculation. This predicted CaMKIl activation will be compared to the measured activation.
Aim 4. Neurogranin (Ng) is an abundant postsynaptic protein that binds CaM and may be important in controlling the CaM that is available to activate CaMKIl. The effects of Ng knockout will be studied.

Public Health Relevance

The proposed research is relevant to addiction, which involves persistent changes in synaptic strength. The role of CaMKIl in the persistence of synaptic strength has recently been demonstrated;notably biochemical attack of CaMKIl has reversed synaptic strength. There is therefore the possiblity that agents that attack CaMKIl can be used to reverse the synaptic changes that underlie addiction.

National Institute of Health (NIH)
National Institute on Drug Abuse (NIDA)
Research Project (R01)
Project #
Application #
Study Section
Special Emphasis Panel (ZRG1-IFCN-B (50))
Program Officer
Sorensen, Roger
Project Start
Project End
Budget Start
Budget End
Support Year
Fiscal Year
Total Cost
Indirect Cost
Brandeis University
Schools of Arts and Sciences
United States
Zip Code
Kabakov, Anatoli Y; Lisman, John E (2015) Catalytically Dead ?CaMKII K42M Mutant Acts as a Dominant Negative in the Control of Synaptic Strength. PLoS One 10:e0123718
Lisman, John; Raghavachari, Sridhar (2015) Biochemical principles underlying the stable maintenance of LTP by the CaMKII/NMDAR complex. Brain Res 1621:51-61
Sanders, Honi; Kolterman, Brian E; Shusterman, Roman et al. (2014) A network that performs brute-force conversion of a temporal sequence to a spatial pattern: relevance to odor recognition. Front Comput Neurosci 8:108
Rennó-Costa, César; Lisman, John E; Verschure, Paul F M J (2014) A signature of attractor dynamics in the CA3 region of the hippocampus. PLoS Comput Biol 10:e1003641
MacGillavry, Harold D; Song, Yu; Raghavachari, Sridhar et al. (2013) Nanoscale scaffolding domains within the postsynaptic density concentrate synaptic AMPA receptors. Neuron 78:615-22
Sanhueza, Magdalena; Lisman, John (2013) The CaMKII/NMDAR complex as a molecular memory. Mol Brain 6:10
Lisman, John E; Jensen, Ole (2013) The ?-? neural code. Neuron 77:1002-16
Kim, Il Hwan; Racz, Bence; Wang, Hong et al. (2013) Disruption of Arp2/3 results in asymmetric structural plasticity of dendritic spines and progressive synaptic and behavioral abnormalities. J Neurosci 33:6081-92
Sanders, Honi; Berends, Michiel; Major, Guy et al. (2013) NMDA and GABAB (KIR) conductances: the ""perfect couple"" for bistability. J Neurosci 33:424-9
Otmakhov, Nikolai; Lisman, John (2012) Measuring CaMKII concentration in dendritic spines. J Neurosci Methods 203:106-14

Showing the most recent 10 out of 25 publications